• Asian-Australasian Journal of Animal Sciences
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• v.33 no.7
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• pp.1068-1076
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• 2020

Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa. Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T₀). The diluted specimens were stored at 4℃ for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T₂₄, T₄₈). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system. Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa. Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

• Polymer(Korea)
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• v.38 no.4
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• pp.530-534
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• 2014

Polyethylene (PE) membranes having various porosities are used as microfilters and separators in lithium ion batteries. Membranes having a high porosity are required for use as separators in a large scale lithium ion secondary battery. In this study, BET was examined for use as a new nontoxic diluent for the fabrication of highly porous PE membranes by thermally induced phase separation process. It was confirmed that BET can be used as a new diluent for the fabrication of the PE membranes by exploring upper critical solution temperature type phase behavior of PE mixtures with BET. When the porosity of the membrane prepared from the PE/PO mixture was compared with that prepared from PE/BET mixture, the latter was about 1.8 times higher than the former.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.12
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• pp.1716-1721
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• 2006

The present study was conducted to observe the effect of osmolality of glycerolated TEST-yolk glycerol extenders on post-thawing sperm kinematics of ram spermatozoa of the native Malpura breed maintained in a semi-arid tropical environment. Good quality semen obtained from adult rams was pooled, split and diluted to 1,000 million spermatozoa per ml in complete TEST-yolk-glycerol extenders of 900, 1,200, 1,500 and 1,800 mOsm/kg osmolality. Diluted semen samples were loaded in 0.25 ml straws and cooled down to $-125^{\circ}C$ freezing temperature at the rate of $-25^{\circ}C$ per minute under controlled conditions before plunging into liquid nitrogen for storage. The thawing of straws was performed at $50^{\circ}C$ in a water bath for 10 seconds and sperm kinematics of the frozen-thawed spermatozoa were assessed by a computer-assisted sperm analysis technique. Osmolality of diluent had no significant effect on post-thawing % motility, % rapid, % medium and % slow moving frozen-thawed spermatozoa but significantly (p< 0.05) affected the % linearity and % straightness. The post-thawing % motility and % rapid motile spermatozoa were highest in samples extended in diluent of 1,500 mOsm/kg osmolality and lowest in 900 mOsm/kg. The curvilinear velocity of spermatozoa was significantly (p<0.05) higher for samples extended in 1,800 mOsm/kg, compared to those in 900 and 1,200 mOsm/kg, but the effect was not significantly different to those extended in diluent of 1,500 mOsm/kg osmolality. The study indicated that ram spermatozoa could tolerate a wide osmolality range for dilution in the complete TEST-yolk-glycerol extender for their cryosurvival. The highest recovery of motile spermatozoa following thawing was achieved in samples extended in the TEST-yolk-glycerol diluent of 1,500 mOsm/kg osmolality.

• Fisheries and Aquatic Sciences
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• v.24 no.2
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• pp.63-77
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• 2021

The aim of this research was to investigate different factors, including cryoprotective agents (CPAs), diluents, dilution ratios, equilibrium times, freezing rates, and thawing methods to optimize cryopreservation protocols for large yellow croaker (Larimichthys crocea). The parameters evaluated were sperm motility, sperm activity index (SAI), survival rate, and DNA damage. Different types of CPAs, such as dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), methanol, and glycerol, were tested for sperm preservation. The highest motility, SAI, and survival rate were observed when EG was used. Different diluents such as Stein's solution, Hank's balanced salt solution, marine fish Ringer's solution, artificial seminal plasma (ASP) of small yellow croaker, and Cortland solution were investigated. The highest post-thaw motility was observed upon using ASP as the diluent. Different concentrations of EG were then mixed with ASP to identify the optimal EG concentration. Experimental results showed that the motility (70.33 ± 1.20%), SAI (5), and survival rate (78.30 ± 0.42%) of post-thaw sperm were optimum when 10% EG and ASP were used as the CPA and diluent of cryopreservation, respectively. Post-thaw sperm motility was high at equilibration times below 150 s and at an optimum dilution ratio of 1:1 (sperm: CPA + diluent) and was not significantly different compared with fresh sperm motility. The freezing rate was found to be slow below -10℃/min. The thawing temperature of 45℃ was identified as ideal. The percentage of tail DNA in post-thaw sperm at 10% EG and ASP was also investigated and was found to have more significant DNA damage than that in fresh sperm but significantly lower damage than that in post-thaw sperm at EG concentrations of 5%, 15%, and 20% (p < 0.05). The cryopreservation protocols obtained in this study will be useful in large yellow croaker hatcheries.

• Fisheries and Aquatic Sciences
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• v.5 no.1
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• pp.64-74
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• 2002

The proper conditions for cold storage of milt in four species of flatfish were investigated. Various diluents were tested for the preservation of marbled sole, brown sole, starry flounder and olive flounder milt at $1\pm0.5^{\circ}C$ for 7 days. The most effective diluent was $1\%$ NaCl for marbled sole and Stein's extender for brown sole, starry flounder and olive flounder. After 7 days of storage in different dilution ratios (milt: diluent), marbled sole milt diluted with 1:3-10 and brown sole, starry flounder and olive flounder milt diluted 1:1-10 maintained good sperm activity index (SAI). SAI was affected by storage volume and presence of antibiotic. SAI was lower after 7 days storage in the volume of 1.5 mL than in 0.5 or 1.0 mL. SAI after 29 days storage was markedly enhanced by addition of antibiotic in marbled sole (400 ppm gentamicin, 400 ppm neomycin, 200-400 ppm streptomycin), brown sole (600 ppm gentamicin), starry flounder (1,000 ppm gentamicin) and olive flounder (200-1,000 ppm gentamicin, 200-400 ppm neomycin, 200-1,000 ppm streptomycin).

• Journal of the Korean institute of surface engineering
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• v.42 no.5
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• pp.227-231
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• 2009

The niobium capacitor showed somewhat more unstable characteristics than the commercial tantalum capacitors, but is nonetheless considered applicable as a future substitute for tantalum capacitors. In this study, niobium powder was fabricated by metallothermic reduction process using $K_2NbF_7$ as the raw materials, KCl and KF as the diluents and Na as the reducing agent. The niobium particle size greatly decreased from 0.7um to 0.2 um as the amount of diluent increased. However if a higher surface area of powder is required, more diluents need to be used in the said method in order to produce niobium powder. The niobium powder morphology and particle size are very sensitive to a amount of sodium excess. The particle size of niobium powder increased with a increasing amount of sodium excess. When more diluent and sodium are used, the niobium powder will be contaminated with more impurities such as Fe, Cr, Ni so on.

• Development and Reproduction
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• v.7 no.1
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• pp.9-13
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• 2003

A series of experiments were conducted to compare the effects of various diluents in cold storage on the sea urchin, Hemicentrotus pulcherimus sperm. Various diluents of glucose solutions, artificial sea water(ASW) and 50% ASW were used to store the sperm at 4$^{\circ}C$. The storage effect was evaluated using sperm activity index(SAI), survival rate of sperm and fertilization rate to egg. ASW and 1.2 M glucose were found to be better diluents which maintained high motility and survival rate of sperm f3r a storage period of 30 days. Optimal pH of diluent to store the sperm at 4$^{\circ}C$ is 7.0∼8.0. In order to keep high SAI and survival rate of sperm, addition of 400 ppm neomycin into the diluent revealed the best storage results.

• Applied Chemistry for Engineering
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• v.7 no.4
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• pp.777-786
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• 1996

Uniform sized-porous poly(acrylonitrile-ethylene glycol dimethacrylate) (PAN) microgels were synthesized by seed polymerization using monodisperse polystyrene seed particles. The effect of weight ratios of monomer/seed (M/S) and diluent/monomer (DIM), concentration of crosslinking agent and the type of diluent was investigated on the formation of PAN microgels. The particle size distribution of PAN microgels was found to be monodisperse in that the weight ratio of M/S is less than 50. More porous microgels were formed and the interstices between small sized interior microspheres present inside of PAN microgels were also smaller with increasing crosslinking agent concentration. Well-developed pore structure was found to be in that weight ratio of D/M is 1 and toluene is used as a diluent.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.12
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• pp.1721-1727
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• 2008

Four experiments were conducted to study the following: i) the influence of different concentrations of sucrose (0.15, 0.3 and 0.5 M with osmolality of 308, 500 and 760 mOsm/kg, respectively) in diluents and control diluent (370 mOsm/kg) on intensity of motility and progressive motility of goat sperm without rehydration and freezing step in four incubation periods (0, 0.5, 2 and 4 h after dilution); ii) the influence of gradual dilution (in 3 steps) on improvements in ascertained results of the first experiment; iii) cryoprotective effects of different concentrations of sucrose (0.15, 0.22, 0.29 and 0.37 M with osmolality of 450, 560, 740 and 920 mOsm/kg, respectively) plus 7% glycerol and 20% egg yolk in basic diluent (Tris-Citric acid-Fructose) and iv) the effect of two concentrations of sucrose (0.15 and 0.22 M) with and without glycerol (7%). In experiment 1, we obtained better results for control diluent, 0.15 and 0.3 M sucrose supplemented diluents with 0 and 0.5 h incubation periods. In experiment 2, apart from a slight improvement, similar tendencies to experiment 1 were observed. In experiment 3, we obtained the best result for diluent with 0.22 M sucrose with regard to intensity of motility, progressive motility, live sperm and normal acrosomes ($40{\pm}4%$, $3.1{\pm}0.2$, $37{\pm}4%$ and $37{\pm}4%$, repectively). In experiment 4, we obtained the best result for diluent with 0.22 M sucrose plus 7% glycerol in regard to intensity of motility, progressive motility and live sperm ($39{\pm}3%$, $3.6{\pm}0.4$ and $41{\pm}4%$, respectively). The characteristic normal acrosomes in diluents without glycerol, i.e. diluents with 0.15 and 0.22 M sucrose showed better results ($39{\pm}8$ and $42{\pm}6%$ respectively). With regard to the release of hyaluronidase enzyme there were no significant differences between diluents (p>0.05). The results of the diluents with 0.15 and 0.22 M sucrose without glycerol were slightly lower than those with glycerol ($69{\pm}11$ and $70{\pm}11$ vs. $72{\pm}11$ and $70{\pm}11{\times}120{\times}10^6$ units $ml^{-1}$, respectively). In conclusion, the use of concentrated sucrose solutions showed that goat sperm can tolerate osmolality up to 560 mOsm (0.22 M) in the freezing period. In addition, glycerol proved to be a necessary cryoprotective agent in the cryopreservation of goat sperm, particularly for intensity of motility, progressive motility and live sperm.

• Journal of the Korean Ceramic Society
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• v.41 no.3
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• pp.235-242
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• 2004

Si, NH$_4$Cl, NaN$_3$, NaCl, $N_2$ were used as raw materials for preparation of $\alpha$-Si$_3$N$_4$ powder. NH$_4$Cl and NaN$_3$ were used as additives, and NaCl was used as a diluent. Initial $N_2$ gas pressure in the SHS reactor was 60 atm. In preparation of $\alpha$-Si$_3$N$_4$, the reactivity and the properties of the products were examined with the various kinds of additives and the content of diluent. At first, the optimum reaction system for the preparation of $\alpha$-Si$_3$N$_4$ is examined and then the optimum composition was examined in the optimum reaction system. The optimum reaction system was Si-$N_2$-additive(NH$_4$Cl+NaN$_3$)-diluent(NaCl) and the optimum composition was 38 wt%Si+50 wt%(NH$_4$Cl+NaN$_3$)+12 wt%NaCl. The maximum fraction of $\alpha$-phase of Si$_3$N$_4$ produced in this condition was 96.5 wt% and the shape of the $\alpha$-Si$_3$N$_4$ produced in this condition was an irregular fiber with a length of 10 ${\mu}{\textrm}{m}$ and a diameter of 1 ${\mu}{\textrm}{m}$.